Directed differentiation of hPSCs through a simplified lateral plate mesoderm protocol for generation of articular cartilage progenitors

Christopher A. Smith; Paul A. Humphreys; Mark A. Naven; Steven Woods; Fabrizio E. Mancini; Julieta O'Flaherty; Qing Jun Meng; Susan J. Kimber

doi:10.1371/journal.pone.0280024

Directed differentiation of hPSCs through a simplified lateral plate mesoderm protocol for generation of articular cartilage progenitors

Christopher A. Smith^*, Paul A. Humphreys, Mark A. Naven, Steven Woods, Fabrizio E. Mancini, Julieta O'Flaherty, Qing Jun Meng, Susan J. Kimber^*

^*Corresponding author for this work

School of Biochemistry

Research output: Contribution to journal › Article (Academic Journal) › peer-review

5 Citations (Scopus)

Abstract

Developmentally, the articular joints are derived from lateral plate (LP) mesoderm. However, no study has produced both LP derived prechondrocytes and preosteoblasts from human pluripotent stem cells (hPSC) through a common progenitor in a chemically defined manner. Differentiation of hPSCs through the authentic route, via an LP-osteochondral progenitor (OCP), may aid understanding of human cartilage development and the generation of effective cell therapies for osteoarthritis. We refined our existing chondrogenic protocol, incorporating knowledge from development and other studies to produce a LP-OCP from which prechondrocyte- and preosteoblast-like cells can be generated. Results show the formation of an OCP, which can be further driven to prechondrocytes and preosteoblasts. Prechondrocytes cultured in pellets produced cartilage like matrix with lacunae and superficial flattened cells expressing lubricin. Additionally, preosteoblasts were able to generate a mineralised structure. This protocol can therefore be used to investigate further cartilage development and in the development of joint cartilage for potential treatments.

Original language	English
Article number	e0280024
Journal	PLoS ONE
Volume	18
Issue number	1
DOIs	https://doi.org/10.1371/journal.pone.0280024
Publication status	Published - 27 Jan 2023

Bibliographical note

Funding Information:
Arthritis Research UK:Susan J. Kimber R20786; MRC UKRMP:Susan J. Kimber MR/K026666; Rosetrees Trust (Rosetrees):Christopher A. Smith, Susan J. Kimber A1984; UKRI EPSRC Centre for Doctoral training in Regenerative Medicine: Paul A. Humphreys, Mark A Naven, Fabrizio Mancini EP/L014904/1; UKRI Medical Research Council (MRC):Qing-Jun Meng MR/T016744/1; UKRI Medical Research Council (MRC): Qing-Jun Meng MR/P010709/1; MRC UKRI Doctoral Training Programme: Julieta O' Flaherty MR/N013751/11.

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Copyright: © 2023 Smith et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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title = "Directed differentiation of hPSCs through a simplified lateral plate mesoderm protocol for generation of articular cartilage progenitors",

abstract = "Developmentally, the articular joints are derived from lateral plate (LP) mesoderm. However, no study has produced both LP derived prechondrocytes and preosteoblasts from human pluripotent stem cells (hPSC) through a common progenitor in a chemically defined manner. Differentiation of hPSCs through the authentic route, via an LP-osteochondral progenitor (OCP), may aid understanding of human cartilage development and the generation of effective cell therapies for osteoarthritis. We refined our existing chondrogenic protocol, incorporating knowledge from development and other studies to produce a LP-OCP from which prechondrocyte- and preosteoblast-like cells can be generated. Results show the formation of an OCP, which can be further driven to prechondrocytes and preosteoblasts. Prechondrocytes cultured in pellets produced cartilage like matrix with lacunae and superficial flattened cells expressing lubricin. Additionally, preosteoblasts were able to generate a mineralised structure. This protocol can therefore be used to investigate further cartilage development and in the development of joint cartilage for potential treatments.",

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Directed differentiation of hPSCs through a simplified lateral plate mesoderm protocol for generation of articular cartilage progenitors

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